Details of experiments between the fragments O60343~595-604 and P62258~1-255
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: TBC1D4
Protein A fragment: 595-604
Protein A site: 14-3-3_pSer600
Protein A modification: (SEP)600
Protein A sequence: SNSLASEKDY
Protein A construct: SNSLA(pSer)EKDY (crude synthetic peptide)

Protein B protein: YWHAE
Protein B fragment: 1-255
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 2

Protein A protein: TBC1D4
Protein A fragment: 595-604
Protein A site: 14-3-3_pSer600
Protein A modification: (SEP)600
Protein A sequence: SNSLASEKDY
Protein A construct: SNSLA(pSer)EKDY (crude synthetic peptide)

Protein B protein: YWHAE
Protein B fragment: 1-255
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.01
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.